Monitoring of Progesterone and Estrone Fecal Metabolites Throughout Gestation in Ewes

Abstract

The aim of this study was to monitor the progesterone and fecal estrone metabolites throughout gestation in ewes correlating with the serum levels of these steroid hormones. Therefore, fecal and serum samples were collected from 5 weeks before mating and gestation until two weeks postparturition. Serum levels of progesterone and estrone and their fecal metabolites were measured by enzyme immunoassay. Serum and fecal hormonal patterns showed a significant correlation for both hormones (R = 0.8572, P < 0.001 for progesterone and R = 0.5893, P < 0.001 for estrone). The fecal progesterone metabolite levels showed significant increasing values among the three thirds of pregnancies, consistent with the serum levels and with the literature. Additionally, the prepartum peak of estrone in the fecal matrix was identified but without observation in the serum matrix due to the blood collection interval used. Therefore, this study demonstrated the viability of progesterone and estrone monitoring throughout gestation using fecal samples, making noninvasive longitudinal endocrine monitoring throughout gestation possible in this species. ResumoO objetivo deste estudo foi monitorar os níveis de metabólitos fecais de progesterona e estrona ao longo da gestação em ovelhas, correlacionando-os com os níveis séricos desses hormônios esteroides. Assim, amostras de fezes e sangue foram colhidas de cinco ovelhas no período pré-cobertura e durante a gestação, até duas semanas após o parto. Os níveis séricos de progesterona e estrona e de seus metabólitos fecais foram mensurados por enzimaimunoensaio. Os perfis hormonais séricos e fecais apresentaram correlação positiva significativa para os dois hormônios (R = 0,8572, P < 0,001 para progesterona e R = 0,5893, P < 0,001 para estrona). Os níveis de metabólitos fecais de progesterona apresentaram valores significativamente crescente entre os terços da gestação, corroborando com os níveis séricos e com os relatos da literatura. Adicionalmente, foi possível evidenciar o pico pré-parto de estrona na matriz fecal, porém sem registro na matriz sérica, provavelmente devido ao intervalo de coletas aplicado. Deste modo, este estudo demonstrou a AMARAL, R.S. et al.Cienc. anim. bras., Goiânia, v.20, 1-10, e-54208, 2019 viabilidade do monitoramento dos níveis de progesterona e estrona durante a gestação em ovinos utilizando amostras fecais, possibilitando monitoramento endócrino longitudinal não invasivo durante a gestação nessa espécie.

Evaluation of Extraction Methods for Progesterone Metabolite Determination in Buffalo Feces by Immunoassay

“…In Ruminant species such as Larter et al, (1994), Kornmatisuk et al, (2007, Isobe et al, (2005) in cows, Hattab et al, (2000), Arunji, (2008), Ashok, (2011 in buffaloes and Jack et al, (2012) in goats. Extraction of fecal progesterone using 90% methanol in black rhinoceros by Edwards et al, (2014), Chelonians by Umapathy et al, (2015) and by 80% methanol in cattle by Palme et al, (1997), goats by Airin et al, (2020), ewes by Amaral et al, (2019), Tsushima leopard cat by Adachi et al, (2010) as well as in Asian Elephants by Kumar et al, (2014Kumar et al, ( , 2019 and guanacos by Marozzi et al, (2019). This study showed a high FPM (5α-pregnan-3α-ol-20-one) value using 90% methanol as a solvent in the dried fecal sample.…”

“…The steroid hormone metabolite quantified in faeces, commonly used in wildlife studies, as a non-invasive, non-stressing, economical, and animal-saving technique that permits longitudinal studies by allowing frequent sampling of the same individual (Brown et al, 1994;Graham et al, 1995). Thus, developing enzyme immunoassays (EIAs) against Progesterone metabolites would be more accurate and reliable for non-invasive reproductive monitoring method using fecal samples (Brown et al, 1994;Umapathy et al, 2013;Mithileshwari et al, 2016;Budithi et al, 2016, Marozzi et al, 2019, Amaral et al, 2019. In various studies, the organic solvent was used by the various workers for the extraction of steroid hormone metabolites from feces.…”

Evaluation of the use of dry blood spots for progesterone determination in ewes

“…The serum samples and the resuspension of dry spots were subjected to a hormonal extraction protocol with diethyl ether described by Rasmussen et al (1996) and already used in sheep (Amaral et al, 2019). In summary, 300 μl of each sample was added to a glass tube along with 1.5 ml of diethyl ether.…”

“…Hormonal analysis was performed by enzyme immunoassay using a protocol described for several other species, including sheep (GRAHAM et al, 2001;AMARAL et al, 2019). Briefly, 96-well polystyrene microplates were labeled with an anti-progesterone antibody (CL425, Davis University -UC Davis, USA).…”

Reproductive and fertility parameters of ewes from the Colombian tropical highlands

“…For the study of the estrous cycle, it has also been analyzed in addition to progesterone, and estrogens in feces, giving good information about the reproductive behavior in sheep (Amaral et al 2019).…”